Novel Triple Combo Adjuvants (Polyphospazenes, CpG-ODNs, Host Defense peptides) mechanism of Immune-stimulation by induction of Chemokines and Cytokines.

1. NOVEL STRATEGIES FOR
IMMUNIZATION AGAINST
PERTUSSIS AND RSV INFECTIONS
Srinivas Garlapati
Vaccine and Infectious Disease Organization
Saskatoon, Canada

2. A novel adjuvant platform for neonates
Polyphos-
phazenes
Delivery of Antigen
Host
Defense
Peptides
Recruitment
CpG ODN
Immune stimulation
Antigen
Platform technology: Pertussis, RSV, others….

3. Courtesy: Parallel solutions Inc
Generation of microparticles
Andrianov et al, 1998

4. Generation of microparticles
1000X 5000X
OVA CpG Merge
Garlapati et al 2008
Garlapati et al 2010

5. Release Kinetics
Garlapati et al 2010

6. Local microenvironment
Alum SOL
MP
MP

7. Triple Combo Soluble Formulation
Activation of IRF by cytosolic pattern recognition receptors

8. Antigen Presentation Pathway
SOL

9. Triple Combo Microparticle
IL-1β Pathway

10. MPs induce STAT and IRF transcription genes

11. MP induce genes in the NFkβ complex

12. PCEP particle formulation by
Ion-bridging with poly-amines
 Single step process by simple mixing PCEP with
natural poly-amine Chitosan or synthetic poly-
ally-amine hydrochloride (PAH) – FDA approved
NaCl-CaCl2 PAH
Chitosan

13. PCEP- CaCl2
MPs
PCEP- PAH
MPs
PCEP- CHIT
MPs
PCEP particle uptake by DCsPCEP particle uptake by DCs
Garlapati et al manuscript in submission

14. Garlapati et al manuscript in submission

15. RSV Vaccine Development

16. RSV MARKET POTENTIAL

17. Non-Replicating RSV vaccines need TLR Adjuvant
Immune response FI-RSV Reason Read out
Humoral response Low Avidity Non-neutralizing Ab-
Ag complexes,
Complement
Activation,
Immune pathology
ELISA-Urea Wash
reduces Absorbance
High virus titers
High Ab/VNT Ratio
Low IgG2a/IgG1 Ratio
Secretory Antibodies Low IgA Parenteral route Low IgA Abs
Cytokines Pro-inflammatory
IL1β & IL-15
Th2
IL-4, IL-5, IL-13
Chemokines
MIP1β, MCP-1, IL-8
Eosinophila,
Peri-vasculitis
Peri-bronchiolitis
Broncho-constriction
Mucus
CD4+ T cells High Prolonged innate
immune responses,
Suppress CD8+ T cells
CD8+ T-cells Low Defective viral
clearancd
High virus shedding

18. RSV VACCINE PIPELINE
Discovery Pre-Clinical Phase-I Phase-II Phase-III
2020
MEDI-559
MEDI-534 (PIV3)
Novavax VLP F & G
PEV4 (virosome)
Protollin- RSV
RSV287
Nanobio RSV
Sendai Virus vector
Alpha Virus vector
Virosomes P3CSK4
rRSVΔG
DNA electoporation
Adenovirus vector

19. 0 21 44 48 50
Immunize Boost Challenge
Kill 1
Kill 2
Spleens - ELISPOT
Lung-Fragment cultures
BAL
Groups
1.Saline/RSV challenge
2.Ag IN
3.Ag Triple Combo SOL IN
4.Ag Triple Combo MP IN
5.Ag IM
6.Ag Triple Combo SOL IM
7.Ag Triple Combo MP IM
Triple Combo formulation with ΔF protein
Viral Shedding
Lung Cytokines

20. RSV-specific humoral responses in serum.

21. Serum Virus Neutralization TitersVirus Titers

22. Lung Fragment Culture

23. Cytokine ELISPOT of Splenocytes

24. Cytokines in Lung Homogenates Post-RSV Challenge

25. Acknowledgements….Acknowledgements….
Nelson Eng
Gael Auray
Tadele Kiros
Rachelle Buchanan
Stacy Strom
Monika Polewicz
Aleksandra Gracia
Laura Latimer
Marlene Snider
Ravinder Garg
Volker Gerdts
Lorne Babiuk
Andrew Potter
Sylvia v.d. Hurk
Heather Wilson
George Mutwiri
Jim Holmstrom
Hugh Townsend
VIDO
Funding: - B. & M. Gates Foundation
- Canadian Institute of Health Research
- Krembil Foundation
- National Science and Engineering Research Council
- Saskatchewan Health Research Foundation
Animal Care
Barry
Walker
Sherry
Others…